Developed for the Determination of Serum Bilirubin between Oxidase Method and Vanadate Oxidation Method / 现代检验医学杂志

Objective To study reaction principle of bilirubin vanadate oxidation method and bilirubin oxidase method through comparison of the determination results,and discuss similarities and difference between the two methods .Methods 310 ca-ses were measured and analyzed with each method.Abnormal samples were further investigated.Results Fortotal bilirubin, the regression equation obtained wasY=1.065 1X+1.197 2,the correlation coefficientr=0.997 0.For direct bilirubin of a-dults and children greater than 30 days,the regression equation wasY=0.945 9X+0.599 5 and the correlation coefficient r=0.994 4.For neonatal direct bilirubin,the regression equation wasY=0.410 4X+2.756 3 and the correlation coefficient r=0.883 5.The results from vanadate oxidation method were unacceptable for abnormal neonatal serum measurement after serial dilution.Conclusion The overall conclusions were that for the measurement of total bilirubin,and direct bilirubin for adults and children older than 30 days.The correlation between these two methods is in an acceptable range,for measure-ment of neonatal direct bilirubin,the correlation between thesetwo methods was not acceptable.It is not recommended to measure neonatal direct bilirubin by vanadate oxidation method.

Determination of Tryptophan Based on Inner Filter Effect of FluorescentYttrium Vanadate: Europium Nanoprobes / 分析化学

Yttrium vanadate:europium nanoprobes (YVO4∶Eu NPs) with good fluorescence properties and water solubility were synthesized by solvent thermal method.Due to the overlapping of the excitation spectrum of YVO4∶Eu NPs and the absorption spectrum of tryptophan, fluorescent internal filter effect (IFE) occurred, in which YVO4∶Eu NPs were the fluorophore and tryptophan was the absorber, leading the fluorescence of YVO4∶Eu NPs was quenched.Therefore, a new method for the determination of tryptophan was established by using fluorescent YVO4∶Eu as nanoprobes based on IFE.Some experimental parameters, such as the adding amount of YVO4∶Eu NPs, pH value of the reacting solution, and reacting time, were investigated.Under the optimum reaction conditions, the linear range of the method was 4.0×10-6-4.0×104 mol/L and the detection limit was 1.0×10-6 mol/L (3σ).The content of tryptophan in the soy sauce was determined with the recovery of 95.2% and 97.3%.This method is simple, rapid, sensitive and accurate.

Influence of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin / 国际检验医学杂志

Objective Discuss the interference of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin .Methods Collected 60 examples ,include total bilirubin concentration 20 examples less than 20 μmol/L ,20 examples between 150-220 μmol/L and 20 examples between 350-410 μmol/L ,add an equal volume of various concentrations of cefotiam in each case ,formulated into cefotiam final concentrations of 300 ,150 ,75 mg/L of serum samples as the test group ,add an equal volume of water in each serum samples as the control group ,determine all the samples total bilirubin concentration respectively by vanadate oxidation method and dry chemical method ,compared the interference of cefotiam on determined total bilirubin by two method ,analyze the data by SPSS13 .0 .Results Determined total bilirubin by dry chemical method ,the test group higher than the control group ,the difference was statistically significant(P<0 .05) ,at the same total bilirubin levels ,with cefotiam concentrations decreased ,increased rate of total bilirubin concentration were decreased in the experimental group .Determined total bilirubin by vanadate oxidation method ,when the total bilirubin concentration between 150 -220 μmol/L ,the test group was higher than the control group ,the difference was statistically significant(P<0 .05) .Conclusion Interference of injection cefotiam on determined to‐tal bilirubin by dry chemical method is strong ,and with the drug concentration increased ,effect is more obvious ,but determination of total bilirubin by vanadate oxidation method has almost no effect .

Inhibitory and stimulating effect of magnesium on vanadate-induced lipid peroxidation under in vitro conditions.

The behaviour of Mg related to vanadium(V)-induced lipid peroxidation (LPO) under in vitro conditions was examined. The studies performed on the liver supernatants (LS) obtained from control, sodium metavanadate-intoxicated, and sodium metavanadate-magnesium sulphate-administered male Wistar rats revealed and confirmed the pro-oxidative potential of V. Simultaneously, they indicated that the improved Mg status may be one of the mechanisms by which the treatment with this element may contribute to reduction of oxidative stress under the conditions of vanadate exposure. On the other hand, the results confirmed that Mg may also stimulate LPO and demonstrated that the incubation conditions and the experimental treatment of the rats from which the liver supernatants were obtained affect the intensity of the examined free radical process.

Vanadium supplementation reduces high fat diet induced hyperlipidemia, hyper insulinemia & hyperglycemia in rabbits.

Vanadium is considered as an essential trace element in some animals. In human’s classification of vanadium as an essential nutrient is still a topic of debate among various research groups. Nutritionally vanadium is thought to be a cofactor in various enzymatic reactions. Increased levels of insulin in blood (hyperinsulinemia) associated with type-2 diabetes mellitus. Increase intake of fat induces hyperinsulinemia which may leads development of type-2 diabetes mellitus. The present study aimed to know the effect of vanadium supplementation on high fat diet induced hyperlipidemia, hyperinsulinemia and hyperglycemia. In this study New Zealand white breed male rabbits divided into three groups. Group-I: rabbits fed with standard diet Group-II: fed with group-I diet and egg yolk, Group-III: rabbits fed with group-II diet and supplemented with 0.5mg/kg of elemental vanadium as sodium meta vanadate. Total cholesterol, LDL-cholesterol and Triglycerides were significantly decreased in G-III when compared to G-II after the experiment. HDL-cholesterol levels are similar in G-II & G-III. Plasma glucose and insulin levels were significantly decreases in G-III than G-II. The present study shows the antidiabetic and antilipidemic role of vanadium in the experimental rabbits. Supplementation of vanadium may prevents hyperglycemia and cardiovascular risk factors like, insulin resistance and hyperlipidemia in diabetes mellitus.

Effects of vanadate and potassium depletion on renal H, K-ATPase protein expression

Background: Vanadate (V) inhibits while potassium (K) depletion stimulates collecting tubule H, K-ATPase activity. In the presence of V, K depletion could not restore the decreased H,K-ATPase activity. The effects of V and K depletion on renal H,K-ATPase protein expression might explain such observation.Objective: To examine the effects of V and K depletion on renal H,K-ATPase protein expression. Methods: Rats treated with normal saline solution (NSS) or V (5 mg/kg body weight) received either normal potassium (NK) or low potassium (LK) diet for 10 days. Protein expressions of renal H,K-ATPase \α₁ and \α₂ isoforms were determined by immunohistochemistry.Results: Both NK and LK animals treated with V had significantly increased vanadium levels in serum, urine, and renal tissues. LK diet caused hypokalemia. Animals treated with LK and V showed progressive hypokalemia. LK stimulated renal H,K-ATPase \α₁ protein expression in both cortex and medulla but enhanced H,K-ATPase \α₂ protein expression only in the cortex. Vanadate did not affect H,K-ATPase \α₁ protein expression in both NK and LK groups. Vanadate unaltered H,K-ATPase \α₂ protein expression in NK animals but could attenuate the increased expression in LK group.Conclusion: The magnitude of direct inhibitory effect of V on renal H,K-ATPase activity with small suppressive effect on protein expression is greater than the stimulatory effect of K depletion on H,K-ATPase activity and protein expression.

Characterization of the 90 kDa heat shock protein (HSP90)-associated ATP/GTPase.

The 90 kDa heat shock protein (HSP90) is an ATP-binding molecular chaperone with an associated ATPase activity having nucleoplasmin and HSP70-binding homology domains and containing Ca-binding EF-hands and a nuclear localization signal. Here we characterize the HSP90-associated ATPase and show that it is (i) a P-type ATPase inhibited by molybdate and vanadate, (ii) able to hydrolyze methylfluorescein phosphate with a 5–6-fold higher affinity, (iii) a 3-times better GTPase than ATPase in the presence of calcium and (iv) HSP27 and F-actin, but not HSP10 can “convert” the HSP90-associated ATPase activity to HSP90 autokinase activity. The HSP90-associated ATP/GTPase may participate in the regulation of complex formation of HSP90 with other proteins, such as F-actin, tubulin and heat shock proteins.

The effects of changes in intracellular Ca2+ activity of osteoblast-like cell on fracture healing / 대한정형외과학회잡지

Bone formation by osteoblast may be closely related to the increase in intracellular Ca2+ activity of osteoblast. In order to study the effects of changes in Ca2+ activity of osteoblast-like cell on fracture healing, we changed intracellular Ca2+ activity of osteoblast-like cells by vanadate and verapamil. And the process of fracture healing was observed after injection of the treatment osteoblast-like cells into the fracture site by hematoxylin-eosin (H-E) stain and bromodeoxyuridine (BrdU) stain. The results were as follow: 1) The most effective range of concentration which could facilitate bone formation was 10-6 to 10-5 M. 2) H-E stain showed more abundant osteoblast and osteoid tissues, more active mitotic division of osteoblast, and earlier appearance of chondroblast and chondroid tissue, making the maturation of woven bone faster in the vanadate-treated group than in the control group. The opposite was true in the verapamil-treated group compared with the control group. 3) BrdU labeling index showed more active osteoblastic proliferation in the vanadate-treated than in the control group. The opposite was observed in the verapamil-treated group compared with the control group. From these results, the fracture healing appears to be facilitated and decelerated by vanadate which apparently increase intracellular Ca2+ activity of osteoblast and verapamil which decreases it, repectively. Therefore the change of intracellular Ca2+ activity of osteoblast can be considered to be one of fracture healing mechanisms and expected to be applied for clinical purpose.

Enhanced Radiosensitivity of Tumor Cells Treated with Vanadate in Vitro / 대한치료방사선과학회지

Intracellular ions which have a major role in cellular function have been reported to affect repair of radiation damage. Recently it has been reported that ouabain sensitizes A549 tumor cells hut not CCL-120 normal cells to radiation. Ouabain inhibits the Na+-K+-pump rapidly thus it increases intracellular Na concentration. Vanadate which is distributed extensively in almost all living organisms in known to be a Na+-K+-ATPase inhibitors. This study was performed to see any change in radiosensitivity of tumor cell by vanadate and any role of Na+-K+-ATPase in radiosensitization. Experiments have been carried out by pretreatment with vanadate in human cell line(A549, JMG) and mouse cell line(L1210, spleen). For the cell survival MTT assay was performed for A549 and JMG cell and trypan blue dye exclusion test for L120, and spleen cells. Measurements of Na+-K+-ATPase activity in control, vanadate treated cell, radiation treated cell (9 Gy for A549 and JMG, 2 Gy for L1201, spleen), and combined 10-6 M vanadate and radiation treated cells were done. The results were summarized as follows. 1. L1210 cell was most radiosensitive, and spleen cell and JMG cell were intermediate, and A549 cell was least radiosensitive. 2. Minimum or cytotoxicity was seen with vanadate below concentration of 10-6 M. 3. In A549 cells there was a little change in radiosensitivity with treatment of vanadate. However radiation sensitization was shown in low dose level of radiation i. E. 2-Gy. In JMG cells no change in radiosensitivity was noted. Both L1210 and spleen cell had radiosensitization but change was greater in tumor cell. 4. Na+-K+-ATPase activity was inhibited significantly in tumor cell by treatment of vanadate. 5. Radiation itself inhibited Na+-K+-ATPase activity of tumor cell with high Na+- K+-ATPase concention. Increase in radiosensitivity by vanadate was closely associated with original Na+-K+-ATPase contents. From the above results vanadate had little cytotoxicity and it sensitized tumor cells to radiation. Inhibitory effect of vanadate on Na+-K+-ATPase activity might be one of the contributing factors for radiosensitization to tumor cells which has greater enzyme activity than that of normal cell. It was suggested vanadate could be used as a potential radiosensitizer for tumor cells.

Stimulation of Cl- secretion by AlF4- and vanadate in T84 cells

We investigated the mechanism of Cl- secretion by fluoroaluminate(AlF4-) and sodium orthovanadate(vanadate) using the human colonic T84 cell line. T84 cell monolayers grown on collagen-coated filters were mounted in Ussing chambers to measure short circuit current(ISC). Serosal addition of AlF4- or vanadate to T84 monolayers produced a sustained increase in ISC. Removal of Ca2+ from the serosal bathing solution partially inhibited AlF4-(-)and vanadate-induced ISC, and readministration of Ca2+ restored AlF4-(-)and vanadate-induced ISC. Carbachol application in the presence of forskolin, AlF4- or vanadate induced a synergistic increase of ISC. Forskolin and vanadate significantly increased cellular cAMP level, while carbachol and AlF4- did not. Carbachol, AlF4- and vanadate significantly increased [Ca2+]i. After Na+ in mucosal bathing solution was replaced with K+, and the mucosal membrane of T84 cell was permeabilized with amphotericin B, AlF4-, vanadate, and carbachol increased K+ conductance, but forskolin did not. After sodium chloride in serosal bathing solution was replaced with sodium gluconate and the serosal membrane was permeabilized with nystatin, forskolin, AlF4-, and vanadate increased Cl- conductance, but carbachol did not. AlF4-(-)induced ISC was remarkably inhibited by the pretreatment of pertussis toxin(2 micrograms/ml) for 2 hours. These results indicate that AlF4- and vanadate can increase Cl- secretion via simultaneous stimulation of Cl- channel and K+ channel in T84 cells. However, the AlF4- action is mostly attributed to stimulation of pertussis toxin-sensitive G-proteins, whereas the vanadate action mostly results from G protein-independent mechanisms.

The effects of vanadium oxide & sodium orthovanadate on murin osteoblast-like (MC3T3-E1) cells / 대한치과교정학회지

Vanadium is an essential trace element but has not been identified with a specific biogical role. To study the direct effects of vanadium on osteoblast, we incubated murin osteoblast-like (MC3T3-E1) cells with various concentration of vanadium oxide & sodium orthovanadate. This study was designed to investigate the effect of vanadium on DNA synthesis, alkaline phosphatase (ALP) activity, cAMP formation responsive to parathormone(PTH) and type I alpha 2 collagen ribonucleic acid (mRNA) level in murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in a -minimal essential medium(alpha -MEM) supplemented with 10% fetal bovine serum (FBS) and then changed to 0.1% FBS with various concentration of vanadium oxide & sodium orthovanadate. Quiescent cultured MC3T3-E1 cells incubated for 24 hours with 2,5,10,15,20 micrometer vanadium oxide incorporated [3H]Thymidine; every concentration showed increases in [3H]Thymidine incorporations dose dependant manner, the greatest response occurred at 20micrometer. Quiescent cultured MC3T3- E1 cells incubated for 3days with 2,5,10,15,20 micrometer vanadium oxide, for 2 days with sodium orthovanadate and alkaline phosphatase was assayed with disodium phenyl phosphate as substrate. Vanadium oxide increased the alkaline phosphatase content in MC3T3- E1 cells at 2 micrometer & 6micrometer; the greatest response occurred at 2micrometer. But decreased at other content. sodium orthovanadate increased alkaline phosphatase content in MC3T3-E1 cells at all concentration ; the greatest response occurred at 4micrometer. Quiescent cultured MC3T3- E1 cells incubated for 3days with 5,10micrometer vanadium oxide, with 5,8micrometer sodium orthovanadate and cAMP formation was measured by Radioimmunoassay(RIA). Vanadium oxide & sodium orthovanadate showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. Quiescent cultured MC3T3-E1 cells incubated for 24hours with 10,20micrometer vanadium oxide, with 5,10micrometer sodium orthovanadate and Type I alpha2 collagen ribonucleic acid (mRNA) expression was studied by Northern blot analysis. Northern blot analysis of vanadium oxide treated cells showed decreasing effects 0& sodium orthovanadate revealed increasing effects in type I &2 collagen ribonucleic acid (mRNA) level.

Effects of sodium vanadate on contractility of vascular smooth muscle

The present study was intended to examine the effect of sodium vanadate on contractility of vascular smooth muscle. Aortic ring preparations were made from the rabbit thoracic aorta and endothelial cells were removed from the ring. The contractility of the aortic ring was measured under various conditions. The results were summarized as follows; 1) Sodium vanadate induced contraction of vascular smooth muscle in a dose-dependent fashion. 2) The contractile effects were not blocked by treatments with adrenergic blocking agent(phentolamine) and indomethacin, indicating the direct action of the drug on vascular smooth muscle. 3) In the presence of ouabain, Na(+)-K(+)-ATPase inhibitor, sodium vanadate still increased the contractility of vascular smooth muscle. 4) Treatment with 4.4'-diisothiocyanostilbene-2.2'-disulfonic acid(DIDS) blocked completely the contractile effects of sodium vanadate. 5) In the presence of verapamil, lanthanum and ryanodine, the contractility of the vascular smooth muscle by sodium vanadate was decreased. From the above results. it was suggested that sodium vanadate acts directly on vascular smooth muscle and causes contraction. It was probably due to inhibition of Ca(++)-ATPase in plasma membrane as well as increasing the release of Ca(++) from sarcoplasmic reticulum and Ca(++) influx across the plasma membrane, but not inhibition of Na(+)-K(+)-ATPase.

Effect of Ammoniummeta Vanadate on Tissue Structure of Jejunum in Rats / 环境与健康杂志

Objective To study the influence of ammoniummeta vanadate on the histological structure of jejunum of rats. Methods One hundred and twelve healthy and clean SD rats were randomly divided into the high dosage (60 mg/L),the moderate dosage(40 mg/L),the low dosage(20 mg/L) and the control group(distilled water),28 in each group,the males and females in the same number,the administration was conducted through drinking water,for 8 consecutive weeks. In the 4th and 8th weeks,the length of jejunum villus,the depth of jejunum gland and the thickness of jejunum wall were measured,the ratio of villus length and gland depth were counted and the pathological examination was done. Results With the increase of ammoniummeta vanadate,the weight of SD rats,the jejunum villus length,the depth of the jejunum gland,the wall thickness had showed a downward trend,the ratio of villus length and gland depth were significantly lower (P

Effects of vanadate on vascular contractility and membrane potential in the rabbit aorta

Isolated rabbit aortic ring with intact endothelial cell preparations precontracted with NE (10(-7) M) were relaxed by vanadate in a dose dependent manner (from 0.2 to 2 mM). Application of vanadate and ACh during the tonic phase of high K+(100 mM)-induced contraction showed a slight relaxation in contrast to that in NE-induced contraction, but sodium nitroprusside (10 microM) more effectively relaxed the aortic ring preparations in high K+ contraction than that of vanadate. Vanadate-induced relaxation in NE-contracted aortic rings was reversed by application of BaCl2 (50 microM) or glibenclamide (10 microM). Furthermore, Vanadate hyperpolarized membrane potential of smooth muscle cells in endothelium-intact aortic strips and this effect was abolished by application of glibenclamide. The above results suggest that vanadate release EDHF (Endothelium-Derived Hyperpolarizing Factor), in addition to EDRF (Endothelium-Derived Relaxing Factor) from endothelial cell. This EDHF hyperpolarize the smooth muscle cell membrane potential via opening of the ATP-sensitive K+ channel and close a voltage dependent Ca++ channel. So it is suggested that the vanadate-induced relaxation of rabbit thoracic aortic rings may be due to the combined effects of EDRF and EDHF.

Effects of Vanadate on the Contractility of Vascular Smooth Muscle

Vanadate is a trace element in animal tissues and has been known to inhibit NA(+)-K(+) ATPase in various tissues including skeletal and cardiac muscles and smooth muscles. Vanadate shows contractile actions on various types of smooth muscles. Prolonged dietary administration of vanadate has been shown to cause arterial hypertension, increased peripheral resistance, and a marked reduction of coronary, visceral and renal blood flow.In isolated vascular smooth muscle of aorta, application of vanadate caused contraction. These studies have been conducted the preparation of vascular smooth muscles from which endothelial cell were removed. It has been reported that endothelial cell releases relaxing factor(s) (endothelium-derived relaxing factor, EDRF) in response to acetylcholine and a number of other stimuli and also produces vasoconstrictor substances (endothelium-derived contracting factor, EDCF). The aim of this present experiment is to elucidate whether vascular response of isolated rabbit aorta induced by vanadate are endothelium dependent or not. The result obtained were summarized as follows ; 1) When endothelium was intact, vanadate induced vascular relaxation of aorta precontracted with norepinephrine. But K+ induced contraction was augmented by vanadate in the aorta with or without endothelium. Whereas relaxation produced by vanadate precontracted with angiotensin II was endothelium-independent. 2) Hemoglobin, methylene blue, hydroquinone, and verapamil inhibited vanadate-induced vascular relaxation. But indomethacin and quinacrine had no effect on vanadate induced vascular relaxation. From the above results, it is speculated the vanadate act on endothelium, modifies the synthesis or release of endothelium-dependent relaxing factor and thus changes the contractile responses to norepinephrine in rabbit aorta.

Optimization of sodium orthovanadate to treat streptozotocin-induced diabetic rats.

Oral administration of sodium orthovanadate restored blood glucose to normal levels in streptozotocin-induced diabetic rats. To establish the safety dose and to evaluate the side effects of over dose, if any, different doses of vanadium were used in the present study. Low concentrations of vanadium (0·1 and 0·3 mg/ml in drinking water) restored blood glucose, urea, cholesterol and the status of liver pathophysiological enzymes to normal levels in experimental rats. High vanadate treatment proved to be toxic not only to diabetic but also to normal rats as evidenced from the observations on the blood urea, plasma and liver glutamate oxaloacetate transaminase and glutamate pyruvate transaminase. Low vanadate treatment restored body homeostasis of diabetic rats and was found to be nontoxic to normals.

Effects of Vanadate on Cellular Ca2+ Movements in Guinea Pig Papillary Muscles

The effects of vanadate on cellular Ca2+ movements across the sarcolemma of cardiac muscle cells were investigated by measuring the intracellular and extracellular Ca2+ activities of guinea pig papillary muscle with Ca2+-selective electrodes. During the rest period following a steady-state of 2 contractions per second the extracellular Ca2+ concentration was increased over the basal level within a minute. During the rest period Ca2+ was transported across the sarcolemma into the extracellular space. Vanadate decreased the change in extracellular Ca2+ concentration during the rest period implying that the Ca2+ efflux across the sarcolemma was decreased by vanadate. Vanadate increased intracellular Ca2+ activities significantly (from 1.9 X 10(-7) M to 10(-6)M) resulting in an increase in resting tension. These results suggest that vanadate decreases Ca2+ efflux from the cells into the extracellular space by blocking Ca2+ transport across the sarcolemma, possibly blocking the Na+-Ca2+ exchange transport.

EFFECT OF SODIUM VANADATE ON INSULIN RECEPTOR BINDING AND GLUCOSE TRANSFER IN STREPTOZOTOCIN-INDUCED DIABETIC RATS / 中华内分泌代谢杂志

After oral administration of sodium vanadate to the streptozotocin-induced diabetic rats for 12 days, the blood glucose of the diabetic rats declined from 21.5?0.7 mmol/L to near normal level, and the water intake of the rats decreased significantly but the serum insulin level did not exhibit marked difference in comparison with the untreated diabetic control nils. The increased percentage of the specific insulin binding to the adipocyte membrane decreased and approached normal range, and a significant increase in glucose transport was found in adipose cell membranes from vanadate-treated rats as copared with those from the normal and the untreated diabetic controls. The above-mentioned findings suggest that the vanadate-induced glucose transport in the tissue cells may play a pivotal role in producing insulin-mimetic effect.

THE EFFECT OF THE INTEGRITY OF NEUROFILAMENT ON THE SHAPE AND DISTRIBUTION OF NEMATOLYSOSOME / 解剖学报

Objective To confirm that if the integrality of NF has effect on the shape and distribution of NLY. Methods using SABC monocolon antibody immunohistochemistry,electron microscopy,immunocytochemistry, and enzymocytochemistry to observe the disorder of NF-H and the changes of distribution and shape of NLY after vanadate treated. Results When the integrality of NF was damaged, the proteins of NF gathered towards nuclear accompany with the similar movements of NLY, meanwhile the shape of NLY also changed into round from thread-like shape.Conclusion Through different ways we used, vanadate can over-phosphate NF proteins and destroy the assemble ability. Whether NF has a complete structure is important to the shape and distribution of NLY, which will be changed when the structure of NF is disorganized.